Immunoliposome labeling: a sensitive and specific method for cell surface labeling.
A simple, one-step procedure for fluorescence labeling of cultured cells with high sensitivity and specificity is described. We term this method immunoliposome labeling. Monoclonal IgG antibody was first covalently coupled with palmitic acid. The palmitoyl IgG was then incorporated into unilamellar liposomes (about 100 nm diameter) containing either N-(4-nitrobenzo-2-oxa-1,3-diazole)- or N-(fluorescein isothiocyanyl)-phosphatidylethanolamine by a detergent-dialysis procedure. A monoclonal antibody to the mouse major histocompatibility antigen, H-2k, was tested as a model system. Fluorescent liposomes with covalently coupled anti-H-2k specifically labeled the mouse L-929 cells (H-2k type), but not the A-31 cells (H-2d type). The degree of labeling was quantitated by a microscope photometer. Cells labeled with fluorescent liposomes showed 4--6-fold stronger fluorescence than cells labeled with either fluorescein-conjugated antibody or with primary antibody followed by fluorescein-conjugated secondary antibody (indirect immunofluorescence). Since different types of label (fluorescent, radioactive, etc.) can be incorporated into liposomes, this specific and sensitive method is potentially very versatile.
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000378:metabolism; D000409:Alanine / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D002462:Cell Membrane / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes; D006041:Goats; D006183:H-2 Antigens; D007074:Immunoglobulin G; D007158:Immunologic Techniques; D008081:Liposomes / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D010714:Phosphatidylethanolamines / Q000378:metabolism; D013861:Thiocyanates / Q000378:metabolism
D000906:Antibodies; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006183:H-2 Antigens; D007074:Immunoglobulin G; D008081:Liposomes; D010714:Phosphatidylethanolamines; D013861:Thiocyanates; C016010:nitrobenzoxadiazolealanine; D009327:4-Chloro-7-nitrobenzofurazan; D016650:Fluorescein-5-isothiocyanate; D000409:Alanine